Tissue engineering. Part C, Methods最新文献

{"title":"Patient-Derived Lung Cancer \"Sandwich Cultures\" with a Preserved Tumor Microenvironment.","authors":"Hailong Wang, Thorsten Walles, Cornelia Wiese-Rischke","doi":"10.1089/ten.TEC.2023.0199","DOIUrl":"10.1089/ten.TEC.2023.0199","url":null,"abstract":"<p><p>In the past, different spheroid-, organotypic-, and three-dimensional (3D) bioprinting lung cancer models were established for <i>in vitro</i> drug testing and personalized medicine. These tissue models cannot depict the tumor microenvironment (TME) and, therefore, research addressing tumor cell-TME interactions is limited. To overcome this hurdle, we applied patient-derived lung tumor samples to establish new <i>in vitro</i> models. To analyze the tissue model properties, we established two-dimensional (2D) and 3D coculture tissue models exposed to static and dynamic culture conditions that afforded tissue culture for up to 28 days. Our tissue models were characterized by hematoxylin eosin staining, M30 enzyme-linked immunosorbent assay, and immunofluorescence staining against specific lung cancer markers (TTF-1 and p40/p63), cancer-associated fibroblast (CAF) markers (α-SMA and MCT4), and fibronectin (FN). The 3D models were generated with higher success rate than the corresponding 2D model. The cell density of the static 3D model increased from 21 to 28 days, whereas the apoptosis decreased. The dynamic 3D model possessed an even higher cell density than the static 3D model. We identified lung cancer cells, CAFs, and FN. Therefore, a novel <i>in vitro</i> 3D lung cancer model was established, which simulated the TME for 28 days and possessed a structural complexity.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"27-37"},"PeriodicalIF":2.7,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10818046/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138806034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Kidney Mesenchymal Stem Cell Differentiation: Effect of Scaffold and Basic Fibroblast Growth Factor.","authors":"Amirhesam Keshavarz Zarjani, Darioush Bijan Nejad, Niloofar Neisi, Mahin Taheri Moghadam, Esrafil Mansouri","doi":"10.1089/ten.TEC.2024.0066","DOIUrl":"10.1089/ten.TEC.2024.0066","url":null,"abstract":"<p><p><b><i>Background:</i></b> Chronic kidney disease (CKD) poses a global health challenge, and it needs alternative therapeutic approaches for patients with end-stage renal disease (ESRD). Although organ transplantation is effective, it faces challenges such as declining quality of life, immunological responses, transplant rejection, and donor shortages. Tissue engineering, by using suitable scaffolds, cells, and growth factors, emerges as a promising treatment option for kidney regeneration. <b><i>Experiment:</i></b> We precisely decellularized scaffold, derived from rat kidneys while maintaining its native three-dimensional (3D) architecture. The efficiency of decellularization was evaluated through histological examinations, including hematoxylin and eosin, periodic acid-Schiff, and DAPI staining, as well as scanning electron microscopy. The scaffolds were then recellularized with kidney mesenchymal stem cells (kMSCs), and their adhesion, proliferation, and differentiation were assessed over 1, 2, and 3 weeks. The expression of specific renal markers, including <i>Wt-1</i>, <i>ZO-1</i>, <i>AQP-1</i>, and <i>ANG-1</i>, was examined through quantitative reverse transcription-polymerase chain reaction (qRT-PCR) in monolayer and 3D cultures. <b><i>Results:</i></b> The infiltration rate of cells into the scaffold increased in a time-dependent manner, and the expression of specific renal markers significantly increased, demonstrating successful differentiation of kMSCs within the scaffold. The application of basic fibroblast growth factor (bFGF) could intensify the expression of kidney-specific genes. <b><i>Conclusions:</i></b> The study highlighted the importance of preserving the 3D architecture of the scaffold during decellularization to achieve optimal cellular responses. Moreover, the capacity of mesenchymal stem cells in recellularized scaffolds facilitated tissue regeneration.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"239-247"},"PeriodicalIF":2.7,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140331990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Young TSPC-Derived Exosomal circPVT1 Ameliorates Aging-Impaired Cell Function via SIRT1/NF-κB.","authors":"Weifeng Han, Dongqiang Gu, Xiaoya Li, Hongguang Chen, Xu Tao, Lei Chen","doi":"10.1089/ten.TEC.2024.0057","DOIUrl":"10.1089/ten.TEC.2024.0057","url":null,"abstract":"<p><p>Tendon stem/progenitor cell (TSPC) senescence is often associated with age-dependent tendon diseases and greatly reduces the capacities for tendon repair and replacement. Exosomes contain bioactive molecules and have been increasingly used in regenerative medicine. In the present study, we demonstrated the antiaging effects of young exosomes from circPVT1-overexpressing TSPCs at early passages (circPVT1-exo). These exosomes attenuated the phenotypes of aged TSPCs at late passages (L-TSPCs) by enhancing self-renewal and proliferation abilities, suppressing cell senescence, maintaining their tenogenic capacity, and weakening their osteogenic differentiation. Mechanistically, circPVT1-exo inhibited the NF-κB pathway and increased SIRT1 expression in L-TSPCs. Knockdown of SIRT1 reversed these effects as evidenced by increased senescence, decreased proliferation, and tenogenic differentiation. These results suggest that circPVT1-exo may ameliorate aging-impaired TSPC function by modulating the SIRT1/NF-κB pathway, suggesting that circPVT1-exo has therapeutic potential for age-related diseases.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"248-254"},"PeriodicalIF":2.7,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141260487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiphoton Imaging of Maturation in Tissue Engineering.","authors":"Maximilian P Werner, Vytautas Kučikas, Kirsten Voß, Dirk Abel, Stefan Jockenhoevel, Marc A M J van Zandvoort, Thomas Schmitz-Rode","doi":"10.1089/ten.TEC.2023.0141","DOIUrl":"10.1089/ten.TEC.2023.0141","url":null,"abstract":"<p><p>Donor cell-specific tissue-engineered (TE) implants are a promising therapy for personalized treatment of cardiovascular diseases, but current development protocols lack a stable longitudinal assessment of tissue development at subcellular resolution. As a first step toward such an assessment approach, in this study we establish a generalized labeling and imaging protocol to obtain quantified maturation parameters of TE constructs in three dimensions (3D) without the need of histological slicing, thus leaving the tissue intact. Focusing on intracellular matrix (ICM) and extracellular matrix (ECM) networks, multiphoton laser scanning microscopy (MPLSM) was used to investigate TE patches of different conditioning durations of up to 21 days. We show here that with a straightforward labeling procedure of whole-mount samples (so without slicing into thin histological sections), followed by an easy-to-use multiphoton imaging process, we obtained high-quality images of the tissue in 3D at various time points during development. The stacks of images could then be further analyzed to visualize and quantify the volume of cell coverage as well as the volume fraction and network of structural proteins. We showed that collagen and alpha-smooth muscle actin (α-SMA) volume fractions increased as normalized to full tissue volume and proportional to the cell count, with a converging trend to the final density of (4.0% ± 0.6%) and (7.6% ± 0.7%), respectively. The image analysis of ICM and ECM revealed a developing and widely branched interconnected matrix. We are currently working on the second step, that is, to integrate MPLSM endoscopy into a dynamic bioreactor system to monitor the maturation of intact TE constructs over time, thus without the need to take them out.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"38-48"},"PeriodicalIF":3.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138805975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Biocompatible Nanofibers Modified by Plasma for Osteoblast Growth Differentiation.","authors":"Quan Liu, Zhongyao Hu, Wendan Cheng, Qinghua Xu, Zhengwei Wu","doi":"10.1089/ten.TEC.2024.0099","DOIUrl":"10.1089/ten.TEC.2024.0099","url":null,"abstract":"<p><p>This work employs nitrogen plasma immersion ion implantation (PIII) to modify electrospinning polylactic acid membranes and immobilizes basic fibroblast growth factors (bFGF) by forming crosslinking bonds. The study investigates the modified membranes' surface characteristics and the stimulatory effects of crosslinked bFGF polylactic acid membranes on osteoblast and fibroblast proliferation. The PIII process occurs under low vacuum conditions and is controlled by processing time and power pulse width. The experimental results indicate that, within a 400-second N₂-PIII treatment, the spun fibers remain undamaged, demonstrating an increase in hydrophilicity (from 117° to 38°/36°) and nitrogen content (from 0% to 7.54%/8.05%). X-ray photoelectron spectroscopy analysis suggests the formation of a C-N-C=O crosslinked bond. Cell culture and activity assessments indicate that the PIII-treated and crosslinked bFGF film exhibits significantly higher cell growth activity (<i>p</i> < 0.05) than the untreated group. These intergroup differences are attributed to the surface crosslinking bond content. In osteogenic induction, the results for each day show that the treated group performs better. However, the intergroup disparities within the crosslinked bFGF group disappear with prolonged culture time due to the rapid osteogenesis prompted by bFGF. The findings suggest that PIII treatment of electrospinning polylactic acid membranes holds promise in promoting osteogenesis in bone tissue scaffolds.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"268-278"},"PeriodicalIF":2.7,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141260383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Developing a Cell-Microcarrier Tissue-Engineered Product for Muscle Repair Using a Bioreactor System.","authors":"Ana Luísa Cartaxo, Ana Fernandes-Platzgummer, Carlos A V Rodrigues, Ana M Melo, Katja Tecklenburg, Eva Margreiter, Richard M Day, Cláudia L da Silva, Joaquim M S Cabral","doi":"10.1089/ten.TEC.2023.0122","DOIUrl":"10.1089/ten.TEC.2023.0122","url":null,"abstract":"<p><p>Fecal incontinence, although not life-threatening, has a high impact on the economy and patient quality of life. So far, available treatments are based on both surgical and nonsurgical approaches. These can range from changes in diet, to bowel training, or sacral nerve stimulation, but none of which provides a long-term solution. New regenerative medicine-based therapies are emerging, which aim at regenerating the sphincter muscle and restoring continence. Usually, these consist of the administration of a suspension of expanded skeletal-derived muscle cells (SkMDCs) to the damaged site. However, this strategy often results in a reduced cell viability due to the need for cell harvesting from the expansion platform, as well as the non-native use of a cell suspension to deliver the anchorage-dependent cells. In this study, we propose the proof-of-concept for the bioprocessing of a new cell delivery method for the treatment of fecal incontinence, obtained by a scalable two-step process. First, patient-isolated SkMDCs were expanded using planar static culture systems. Second, by using a single-use PBS-MINI Vertical-Wheel^® bioreactor, the expanded SkMDCs were combined with biocompatible and biodegradable (i.e., directly implantable) poly(lactic-<i>co</i>-glycolic acid) microcarriers prepared by thermally induced phase separation. This process allowed for up to 80% efficiency of SkMDCs to attach to the microcarriers. Importantly, SkMDCs were viable during all the process and maintained their myogenic features (e.g., expression of the CD56 marker) after adhesion and culture on the microcarriers. When SkMDC-containing microcarriers were placed on a culture dish, cells were able to migrate from the microcarriers onto the culture surface and differentiate into multinucleated myotubes, which highlights their potential to regenerate the damaged sphincter muscle after administration into the patient. Overall, this study proposes an innovative method to attach SkMDCs to biodegradable microcarriers, which can provide a new treatment for fecal incontinence.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"583-595"},"PeriodicalIF":3.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10714258/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41238727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Nerve Damage After an Injury to the Adjacent Soft Tissue: A Pilot Animal Study.","authors":"Nesreen Zoghoul Alsmadi, Curt Deister, Nik Agrawal, Lan Tran, Rasa Zhukauskas, Debbie Neubauer Fischer, Deana Mercer","doi":"10.1089/ten.TEC.2023.0151","DOIUrl":"10.1089/ten.TEC.2023.0151","url":null,"abstract":"<p><p>Traumatic injuries may result in the formation of soft tissue adhesions between peripheral nerves and surrounding soft tissue. These soft tissue adhesions lead to compression and ischemic stress within fascicles due to nonpliability of adhered scar tissue, and nerve tension due to loss of nerve gliding from scar tethering. These changes in the soft tissue bed surrounding the nerve may result in axon degeneration and neuroma-in-continuity. Preclinical models that simulate clinically relevant levels of scar in the nerve environment may be impactful to the development of surgical techniques and treatments to prevent adhesions. This study presents the results of a rodent model with an induced indirect nerve injury by (1) thermal insult to the soft tissue bed surrounding the nerve and (2) air-drying the surrounding soft tissue bed of the nerve. Our findings suggest that inducing an injury of the soft tissue bed results in increased intraneural scar and extraneural adhesions to the nerve compared to a sham procedure. Thermal induced injuries showed more macrophages and changes in nerve health compared to air-dried induced injuries. The changes in the nerves of the induced injury groups, specifically the thermal injury group, may be meaningful for evaluating treatments for nontransected nerve injuries.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"547-557"},"PeriodicalIF":3.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10714259/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41141417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biomimetic Cardiac Fibrotic Model for Antifibrotic Drug Screening.","authors":"Haiyan Li, Yifan Zhu, Zhe Chen, Qiaolin Ma, Ahmed I Abd-Elhamid, Bei Feng, Binbin Sun, Jinglei Wu","doi":"10.1089/ten.TEC.2023.0089","DOIUrl":"10.1089/ten.TEC.2023.0089","url":null,"abstract":"<p><p>Cardiac fibrosis is characterized by pathological proliferation and activation of cardiac fibroblasts to myofibroblasts. Inhibition and reverse of transdifferentiation of cardiac fibroblasts to myofibroblasts is a potential strategy for cardiac fibrosis. Despite substantial progress, more effort is needed to discover effective drugs to improve and reverse cardiac fibrosis. The main reason for the slow development of antifibrotic drugs is that the traditional polystyrene culture platform does not recapitulate the microenvironment where cells reside in tissues. In this study, we propose an <i>in vitro</i> cardiac fibrotic model by seeding electrospun yarn scaffolds with cardiac fibroblasts. Our results show that yarn scaffolds allow three-dimensional growth of cardiac fibroblasts, promote extracellular matrix (ECM) deposition, and induce the transdifferentiation of cardiac fibroblasts to myofibroblasts. Exogenous transforming growth factor-β1 further promotes cardiac fibroblast activation and ECM deposition, which makes it a suitable fibrotic model to predict the antifibrotic potential of drugs. By using this platform, we demonstrate that both Honokiol (HKL) and Pirfenidone (PFD) show potential in antifibrosis to some extent. HKL is more efficient in antifibrosis than PFD as revealed by biochemical composition, gene, and molecular analyses as well as histological and biomechanical analysis. The electrospun yarn scaffold provides a novel platform for constructing <i>in vitro</i> fibrotic models to study cardiac fibrosis and to predict the antifibrotic efficacy of novel drugs.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"558-571"},"PeriodicalIF":3.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10142939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Call for Special Issue Papers:</i> Special Issue for Prof. James Kirkpatrick in Honor of his Achievements in Tissue Engineering/Regenerative Medicine.","authors":"Laura De Laporte, Jeroen van den Beucken","doi":"10.1089/ten.tec.2023.29041.cfp","DOIUrl":"10.1089/ten.tec.2023.29041.cfp","url":null,"abstract":"","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 12","pages":"545-546"},"PeriodicalIF":3.0,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138806129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Organalysis: Multifunctional Image Preprocessing and Analysis Software for Cardiac Organoid Studies.","authors":"Jathin Pranav Singaraju, Adheesh Kadiresan, Rahul Kumar Bhoi, Angello Huerta Gomez, Zhen Ma, Huaxiao Yang","doi":"10.1089/ten.TEC.2023.0150","DOIUrl":"10.1089/ten.TEC.2023.0150","url":null,"abstract":"<p><p>Due to a growing need in visualizing human pluripotent stem cell-derived organoids from recent advancements in the field, an efficient bulk-processing application is necessary to provide preprocessing and image analysis services. In this study, we developed Organalysis, a high-accuracy, multifunctional, and accessible application that meets these needs by providing the functionality of image manipulation and enhancement, organoid area and intensity calculation, fractal analysis, noise removal, and feature importance computation. The image manipulation feature includes brightness and contrast adjustment. The area and intensity calculation computes six values for each image: organoid area, total image area, percentage of the image covered by organoid, the total intensity of organoid, the total intensity of organoid-by-organoid area, and total intensity of organoid by total image area. The fractal analysis function computes the fractal dimension value for each image. The noise removal function removes superfluous marks from the input images, such as bubbles and other unwanted noise. The feature importance function trains a lasso-regularized linear regression machine learning algorithm to identify cardiac growth factors that are the strongest determinants for cell differentiation. The batch processing of this application further builds on existing services like ImageJ to provide a more convenient way to process multiple images. Collectively, the versatility and preciseness of Organalysis demonstrate novelty, since no other current imaging software combines the capability of batch processing and the breadth of feature analysis. Therefore, Organalysis provides unique functions in cardiac organoid research and proves to be invaluable in regenerative medicine.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"572-582"},"PeriodicalIF":2.7,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10714253/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10172385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}